Method for detecting instability of human cell mitotic chromosomes in vitro

Abstract

The invention provides a method for detecting the instability of human cell mitotic chromosomes in vitro. The method comprises the following steps: culturing cells, treating the cells, collecting the cells, concentrating the cells, smearing the cells, fixing the cells, dyeing the cells and sealing the cells; observing mitotic cells by utilizing an optical microscope and carrying out frequency counting on an instability index of the chromosomes. The method is a method for systemically and omnidirectionally detecting the instability of the mitotic chromosomes; compared with other methods, the method has the advantages that (1) an instability phenomenon of a plurality of types of chromosomes can be detected at the same time, and whether a certain compound is a chromosome toxin and / or a mitotic toxin is analyzed; (2) the method has very high sensitivity and can be used for detecting extremely low dosage of genetic toxic substances; (3) a false positive phenomenon caused by artificial reasons can be extremely reduced; (4) the method has the characteristics of simplicity and easiness of operation; the labor force can be effectively saved and the method has a high-throughput application potential; (5) the method has relatively low economic cost and is applicable to development and implementation in most of laboratories.

Description

technical field [0001] The invention relates to the technical field of life sciences, in particular to a method for detecting chromosome instability in human cell mitosis in vitro. Background technique [0002] Mitosis is the most vulnerable phase of the cell cycle as cells are exposed to exogenous or endogenous chemicals that cause chromosomal instability. Chromosomal instability can further lead to genomic instability in cells, which is the underlying cause of many human genetic-environmental diseases, which are associated with birth defects, developmental abnormalities, immune deficiencies and degenerative diseases (such as Alzheimer's disease, Parkinson's disease , hypertension, cardiovascular disease, diabetes, osteoporosis, tumors, etc.) are highly correlated with increased risk. Because there are many indicators of chromosomal instability, most of the detection methods currently used can only target a few of them, and there is a lack of methods that can systematicall...

AI Technical Summary

Problems solved by technology

Although this method can analyze and count most of the chromosomal instability phenomena, the disadvantages of this method are: ① labor-intensive

The chromosome dispersion method has high requirements on the state of chromosome separation, and the number of chromosomes in human cells is large and the size varies greatly. These factors limit the number of effective observations of the chromosome dispersion method, which in turn increases the statistical error and reduces its sensitivity.

② There may be false positive results caused by human factors

However, some other methods for detecting chromosomal instability are faced with the shortcomings of single detection index or high economic cost.

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